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作 者:詹爱军[1] 王新卫[2] 陈书琨[1] 孙洁[1] 陈兵[1] 阮周曦[1] 花群义[1]
机构地区:[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518010 [2]河南农业大学牧医工程学院,河南郑州450002
出 处:《动物医学进展》2010年第2期36-40,共5页Progress In Veterinary Medicine
基 金:科技部"863"计划项目(2006AA10Z445);国家质检总局资助项目(2008IK011)
摘 要:为建立可检测鹿流行性出血病病毒(EHDV)的液相芯片快速检测技术,用DNAStar软件对GenBank中EHDV的VP7基因进行序列分析,设计EHDV特异性探针并用生物素标记,与荧光编码微球偶联后与病毒VP7基因的PCR产物杂交反应,用液相芯片检测仪(Liquichip200)检测荧光信号建立了EHDV快速高通量液相芯片检测方法。检测结果显示,该法具有较好的特异性,不与其他虫媒病病毒反应;检测灵敏度为100个TCID50。建立了快速检测EHDV的液相芯片技术,为进一步搭建EHDV快速高通量检测平台奠定了基础。To develop liquichip method for detecting epizootic haemorrhagic disease virus of deer, the VP7 gene of EHDV published in the GenBank was analyzed by using the software DNAStar 5.0, and specific EHDV probe labeled with biotin was prepared and coupled with fluorescence-coded microspheres, which was used for hybridization reaction to RT-PCR products of EHDV. Liquichip detection method for detecting EHDV was established by using Liquichip 200 to detect fluorescence signal in the reaction system. The results demonstrated that this detection method showed better specificity to EHDV and no reaction with other insect-borne disease viruses. The sensitivity test indicated that the detecting limit for the EHDV could reach to 100 TCID50. These results suggested that the developed Liquichip detection technique provided a foundation for further research on rapidly high throughput detection for EHDV and also for exploration to detect other viruses with the same way.
分 类 号:S858.25[农业科学—临床兽医学] S852.65[农业科学—兽医学]
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